2017 Fiscal Year Annual Research Report
Genetic analysis and investigation of molecular pathogenesis for mitochondrial diseases
| Project/Area Number |
17F17714
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| Research Institution | Saitama Medical University |
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Principal Investigator |
岡崎 康司 埼玉医科大学, 医学部, 客員教授 (80280733)
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| Co-Investigator(Kenkyū-buntansha) |
LIM SZE CHERN 埼玉医科大学, 医学部, 外国人特別研究員
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| Project Period (FY) |
2017-07-26 – 2019-03-31
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| Keywords | Mitochondria / Human genetics / Molecular biology |
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| Outline of Annual Research Achievements |
The aim of my current research is to identify the molecular causes of disease in patients with mitochondrial disease and to discover novel disease genes. I investigated 8 mitochondrial disease patients with candidate mutations in 7 genes, 6 of which have never been linked to human mitochondrial disease. Bioinformatic, molecular genetic and biochemical analyses including Sanger sequencing, DNA cloning, qPCR, SDS-PAGE and BN-PAGE western blotting, respiration rate analysis and OXPHOS enzyme assays had been performed. As a result, three of the genes have been excluded from further analysis due to the lack of evidence for pathogenicity of the candidate mutations. Lentiviral-mediated phenotypic rescue experiments had been performed in fibroblast cell lines from 4 of the patients.
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| Current Status of Research Progress |
Current Status of Research Progress
2: Research has progressed on the whole more than it was originally planned.
Reason
I investigated the molecular causes of disease in 8 patients with early-onset mitochondrial disease. Various bioinformatic, biochemical and molecular genetic analyses were performed. I established a new research collaboration with Dr Diana Stojanovski from University of Melbourne, Australia.
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| Strategy for Future Research Activity |
In FY2018, I will continue my research to confirm the pathogenicity of candidate mutations in 6 patients with mitochondrial disease. I will set up a 2D-PAGE western blotting system in the lab to analyze patient samples. The cell lines I generated from lentiviral-mediated rescue experiments will be analyzed by western blotting and respiration rate analysis.
Furthermore, I plan to improve my computational skill which is fundamental for advanced bioinformatic analysis of next generation sequencing data. I plan to attend the 3rd HVP Variant Effect Prediction Training Course and the Cold Spring Harbor Laboratory Computational Genomics course.
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